Advanced Imaging Core - Abstract The overall goal of the Advanced Imaging Core is to provide DDRCC investigators with sophisticated technologies, methods and training required for successful, high-end cell and tissue-based imaging and analysis, including confocal, multiphoton and super-resolution microscopy, intravital imaging, and automated imaging of histological slides. Specific Aims are: 1) Provide DDRCC investigators with state-of-the-art capabilities in optical microscopy; 2) Develop novel, front-line imaging applications that address the emerging scientific needs of DDRCC members; and 3) Provide expert mentoring, education, training, and consultation on sophisticated optical imaging technologies that enable scientific discovery by DDRCC investigators, as well as the greater MUSC research community. The Core houses the following major microscope systems: 1) Zeiss LSM 880 NLO multiphoton/confocal system equipped with Quasar spectral detection and Fast Airyscan super-resolution capability; 2) Olympus Fluoview FV1200 multiphoton microscope with silicone oil optics for intravital imaging; 3) Olympus Fluoview FV 10i LIV live cell confocal microscope with water immersion optics; 4) Zeiss LSM 510 META confocal microscope; 5) BD BioSciences CARV II disk-scanning confocal microscope for video-rate imaging; 6) Zeiss Axiovert 200M widefield fluorescence microscope; and 7) Perkin-Elmer Vectra Polaris Automated Quantitative Pathology Imaging System. Except for Vectra Polaris which is customized for histological slides, all microscopes are equipped with environmental chambers for temperature and gas phase control to allow non- destructive 3D imaging of living cells, tissues and organisms. Major applications include 1) live cell super- resolution imaging of parameter-indicating fluorophores to monitor ions, electrical potentials, radical generation, pyridine nucleotide reduction, membrane permeability, cell viability (apoptosis and necrosis) and the submicron distribution of fluorescent proteins and other fluorescent reporters; 2) high resolution imaging of tissue sections for immunocytochemistry and fluorescent protein distribution; 3) fluorescence resonance energy transfer (FRET) and DuoLink to characterize and quantify interactions between specific molecules; 4) intravital microscopy to monitor microcirculation, leukocyte margination, mitochondrial polarization and permeability, radical generation, gene expression and other parameters in living animals; and 5) high throughput, quantitative multiplexed imaging of conventionally and immuno-stained clinical and research specimens. Computer workstations provide offline image processing/analysis (Bitstream Imaris, ImageJ FIJI, Metamorph, IPLab and others). Hands-on and didactic training in current and next-generation imaging approaches are provided by seminars, demos and a biennial 1-week Charleston Workshop on Light Microscopy for the Biosciences. The Advanced Imaging Core services promote the success of DDRCC investigator research on digestive and liver disease.